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Introduction: Background: Neff regulatory protein as well as structural proteins such as MPER and V3 of HIV-1 is important targets in vaccine studies. Therefore, in the current study, the Nef-MPER-V3 fusion protein was cloned and expressed in prokaryotic expression system that will be evaluated as a candidate for future vaccine studies. Materials and Methods: The Nef-MPER-V3 sequence was synthesized and inserted into pET-28a(+) vector using Not I/HindIII enzymes. Then, the recombinant construct was expressed in BL21 (DE3) strain of E.coli. For optimization of protein expression, several factors such as time of induction, IPTG concentration and optical density (OD) were evaluated. The recombinant protein was purified on a Ni-NTA agarose column and analyzed by 12% SDS-PAGE. Finally, the fusion protein was confirmed by western blotting. Results: The target gene was successfully cloned into the recombinant vector. The best condition for the expression such as; 2 m IPTG, OD600 = 0.7 and growth 5 hours post-induction were determined and the recombinant protein was purified by affinity chromatography in denaturing condition with urea at a concentration of about 300 μg/ ml. The purified fusion protein migrated as a clear band of around 35 kDa in SDS-PAGE and was detectable using anti-Neff antibody in western blotting. Conclusion: Our results showed that the Nef-MPER-V3 protein was successfully expressed in prokaryotic system and purified protein may provide the antigen for future experiments for immunogenicity evaluation in mice is currently undertaken. [ABSTRACT FROM AUTHOR]

Background and Aims: Many bacteria including Escherichia coli may enter into a viable but non-culturable (VBNC) state under unfavorable stresses, which are unable to be detected by culture-based methods. In this study, the use of Reverse Transcription PCR (RT-PCR) for detection of VBNC state of E. coli O157:H7 was investigated. Materials and Methods: Escherichia coli O157:H7 was inoculated in distilled water and 30 percent salt water at room temperature. The cultivability of bacteria was determined using routine culture and colony counting on MacConkey agar. The RT-PCR of 16S rRNA gene involved direct extraction and purification of RNA, DNase I treatment for removing DNA contamination, cDNA synthesis and electrophoresis of PCR products of cDNA was used to detect viable E. coli O157:H7 under studied treatments and was compared with the results of RT-PCR of 16S rRNA gene of heat- killed bacteria. Results: The cultivability of bacteria was maintained during the study period in distilled water; however, the use of 30% NaCl caused the bacteria to be non-cultivable on day 4. The RTPCR of 16S rRNA showed the positive expression of this gene in cultivable and non-cultivable bacteria during the study period, whereas heat-killed bacteria were negative for this gene, which indicated the efficacy of RT-PCR of 16S rRNA in differentiation of alive from dead bacteria. Conclusions: Escherichia coli O157:H7 entered into the VBNC under 30 percent NaCl which can be associated with serious human health problems. RT-PCR can be used to detect bacteria in the VBNC state. [ABSTRACT FROM AUTHOR]

Application of lysozyme as a natural antimicrobial or preservative agent in food and pharmaceutical industry is well known. The antimicrobial effects of lysozyme can be expanded by conjugation with carbohydrates via Maillard reaction, however this natural reaction is a time consuming process. So in this study a new method microwave heating, was applied to Accelerate the Maillard reaction. Lysozyme was mixed with dextran and heated by microwave irradiation. The glycation extent was determined by SDS-PAGE electrophoresis, cation-exchange chromatography and sugar analysis. SDS-page pattern show that with increasing heating time a high molecular weight conjugates were appeared and maximum glycosylation was occurred after 4 minutes heating. The elution profiles (chromatograms) of the lysozyme–dextran conjugates compared the natural lysozyme showed that the conjugated peak shifted towards the higher molecular weight fraction and overlapped with the non conjugated proteins, indicated that lysozyme was covalently bound to dextran. These results suggest that Maillard reaction rate can be increase using microwave heating. [ABSTRACT FROM AUTHOR]

Background: DNA-based computing is an emerging research aspect that enables the in-vivo computation and decision making with significant correctness. Recent papers show that the expression level of miRNAs are related to the progress status of some diseases such as cancers and DNA computing is introduced as a low cost and concise technique for detection of these biomarkers. In this paper, DNAbased logic gates are implemented in the laboratory to detect the level of miR-21 as the biomarker of cancer. Materials and Methods: At the first, required strands for designing DNA gates are synthesized. Then, double stranded gate is generated in laboratory using a temperature gradient that followed by electrophoresis process. This double strand is the computation engine for detecting the miR-21 biomarker. miR-21 is as input in designed gate. At the end, the expression level of miR-21 is identified by measuring the generated fluorescent. Results: at the first stage, the proposed DNA-based logic gate is evaluated by using the synthesized input strands and then it is experimented on a tumor tissue. Experimental results on synthesized strands show that its detection quality/correctness is 2.5x better than conventional methods. Conclusion: Experimental results on the tumor tissues are successful and are matched with those are extracted from real time PCR results. Also, the results show that this method is significantly more suitable than real time PCR in view of time and cost. [ABSTRACT FROM AUTHOR]

Background and Aims: Brucellosis is an infectious zoonotic disease responsible for reproductive disorders among animals and significant economic losses to the meat and milk supplychain. The purpose of this study was therefore to evaluate and diagnose the species and strains of Brucella isolates among cattles via multiplex polymerase chain reaction. Materials and Methods: In this study, blood samples were taken from 22 cows diagnosed as having brucellosis by PCR test. Blood samples were cultured in BACTEC vials and incubated at 37 degrees Celsius for 5 days following which each sample was cultured on Brucella agar and kept for 3 days under Co2 condition.The 16SrRNA gene was used as the target for the detection of Brucella genus and the bcsp31 gene was used for the detection of Brucella biovars. The PCR products were electrophoresed on %1 agarose and compared with standard strains. Results: The PCR results showed a total of 22 positive samples for Brucella among cattles. Out of the 22 cattle positive samples, 19 were identified as B.abortus (biovars 1, 2, 3, 4, 5) and 3 as B. melitensis (biovars 1, 2, 3). Conclusions: Brucella abortus biovars 1 and 3 and B .melitensis biovar 1 are the most prevalent biovars among cattles in Lorestan province, Iran. Moreover, multiplex PCR method could be used to improve the qualitative level and the Brucella diagnosis speed in laboratories. [ABSTRACT FROM AUTHOR]

Background and Objectives CD40L is a membrane protein and a member of the tumor necrosis factor super family (TNFSF) that plays an important role in transferring cell signaling in innate and adaptive immunity. Since this protein plays its role through clustering the receptors on the target cell surface, it seems that the multimeric form of this molecule can have a stronger effect than the natural trimeric form. So in this study we describe cloning and expression of dodecameric soluble CD40L through self multimerizing protein, surfactant protein D (SP-D), in the HEK293 cell line. Materials and Methods In this experimental study, the SPD-CD40L was designed in silico and cloned in pcDNA3.1(+) plasmid. HEK293 cells were transfected and used as the expression host cells. The expression of SPD-CD40L was determined at the RNA level by RT-PCR method. After the purification of the recombinant proteins by affinity chromatography, the molecular weight of the recombinant protein was determined by SDS-PAGE. To evaluate the specificity of protein, ELISA and Dot Blot were used. Results The results indicated that HEK293 cells were transfected and recombinant protein was expressed 24 and 48 hours after Transfection. Moreover, the results of ELISA and Dot Blot methods represented the specificity of recombinant SPD-CD40L chimeric protein. Conclusions Chimeric protein SPD-CD40L was expressed successfully by pcDNA3.1(+) in eukaryotic HEK293 cell line and the molecular weight of 4-trimeric (dodecameric) protein was shown to be consistent with the expected amount. [ABSTRACT FROM AUTHOR]

Background and Objectives There is considerable phenotypic variation of Beta-thalassemia with common mutations whose understanding would facilitate the prevention of the specific syndromes. The aim of this study was to determine the common Beta-thalassemia mutations in Isfahan and Chaharmahal Bakhtiari province and thier relationship with blood parameters. Materials and Methods In this descriptive study, 10 ml venous blood samples were taken from 321 Beta thalassemia carriers having referred to the private Isfahan Center of Medical Genetics in 2016. For the confirmation of the diagnosis of Beta thalassemia in these patients, the indices of MCV and MCH with Mindary device and HbAl, HbA2, Hbf and RBC with the electrophoresis method were measured, yf, t-test and SPSS 22 were used in analyzing data. Results Mutation Fr36/37 (-T) in the population studied in Isfahan and Chaharmahal Bakhtiari with the frequency rates of 34 (26.35%) and 22 (32.35%) showed the highest in the studied mutations. About 80% of cases were detectable mutations in the beta globin gene in the people with HbA2 > 3.5 and in 100% of cases with MCH < 27 and MCV < 80. Conclusions Beta-thalassemia mutations among Chaharmahal Bakhtiari and Isfahan populations show diversity and wide distribution. Average mutations studied based on blood indices showed a wide variety. The results showed a positive correlation between beta-thalassemia mutations and red blood cells indices which can be effective in fast and efficient screening of these common mutations. [ABSTRACT FROM AUTHOR]

Background: Leishmaniasis is a protozoan parasitic disease and a global health problem. The aim of this study is to diagnose the parasitic infection in humans for epidemiological identification and providing control programs using proprietary co-designed primers in three species of Leishmania. Materials and Methods: 30 common Leishmania isolates were gathered from different centers in Iran. Having been cultured in RPMI-1640 Medium, DNA was extracted and the gene ITS2-rRNA was amplified by PCR. The amplicons were examined by electrophoresis on agarose gel 2%. Also, in FLASH PCR method, a specific probe and florence colour were used to investigae the amplicon existence on sample. Results: The results of the investigations by PCR and FLASH PCR methods show that these methods are sensitive and specific for diagnosis of Leishmania Conclusion: In this study, identification of Leishmania parasite using specific primer pairs was successful and TaqMan could be one of the most sensitive diagnostic methods to identify parasite load for the ITS2 region of Leishmania [ABSTRACT FROM AUTHOR]

Introduction: Nowadays, rising of cancer prevalence has increased the importance of its treatment. Leukemia is the cancer of blood-forming tissues which derives from red and white blood progenitors cells. Prednisolone (Family members of glucocorticoid drugs) with anti-tumor and apoptotic effects has been considered by many researchers. Objective: The aim of this study was the investigation of the effect of prednisolone on K562 cell line. Materials and Methods: First, The K562 cells were cultured. After, by using different concentrations of prednisolone, antitumor properties of this drug on K562 cells was measured by MTT assay after 72 hours. Finally, Hoechst (33342) staining and DNA electrophoresis were used for study of apoptosis. Data analysis was performed using SPSS software and ANOVA test (P<0.05). Results: In this study, anti-tumor effect of prednisolone on K562 leukemia cell line was confirmed. The results also showed that prednisolone, compared with the control group, had cytotoxic activity that is highest in 100μM at 72 hours.Statistical analysis indicated that this drug inhibited the growth of K562 cells (P < 0.05). Conclusion: Based on the results, prednisolone is an effective agent for chronic myeloid leukemia therapy and prevention. We suggest that further studies on the effects of this drug on animal models and human be done. [ABSTRACT FROM AUTHOR]

The genus Salvia (Lamiaceae) comprises over 900 species in the world, with a relatively wide dispersion in the Iran's flora. Until now, about 58 species of the genus have been reported and identified in Iran, in which 17 of them were endemic. In order to study, investigate and evaluate the intraspecific diversity, similarity and dissimilarity among Iranian Salvia officinalis accessions, an experiment was carried out using SDS-PAGE technique. In this study, the seeds from five accessions were collected from gene bank and were evaluated. The seed storage proteins were extracted by buffers and were measured. Phylogenetic relationships were analyzed according to presence and absence of bands on the gel. A dendrogram was prepared using calculation of the accession's similarity index. Mean comparison were done by Tukey's test. The seed protein contents showed significant differences (p≤0.01) among accessions. A total of 39 bands were indicated on the gel. The maximum diversity was detected in the accession No. 2 while, the lowest band's number were recorded with the accessions No. 1 and 5. Based on dendrogram, the accessions were divided into two groups; one includes accession No. 1 and 4 other accessions were located in the second group further classified into two subgroup including accessions No. 2 and 3 in one clade and accessions No. 4 and 5 in the other ones. [ABSTRACT FROM AUTHOR]

Background and purpose: The purpose of this study was to design a recombinant vector pEFGP- N1 containing the full length of HIV-1Vpr gene. To the best of our knowledge, the cloning of Vpr gene in pEGFP_N1 is not previously done. Materials and methods: As a source of Vpr gene the pUC19-Vpr recombinant vector was confirmed by digestion with restriction enzymes BglII and NotI in order to separate the Vpr gene. The specific primers for Vpr gene including the restriction sites of XhoI and KpnI were designed according to the multiple cloning site of pEGFP-N1 and PCR reaction was performed using the pUC 19-Vpr vector as a template. The PCR product was undergone electrophoresis and gel extraction. Digestion reaction was done on both the extracted PCR product and the pEGFP-N1vector. The recombinant pEGFP-N1-Vpr vector was achieved by the ligation reaction using the T4 DNA ligase and it was transformed into the E-coli (DH5α) and propagated. Finally, confirmation was done through the restriction enzyme digestion and PCR amplification. Results: The recombinant vector pUC19-Vpr was confirmed using the restriction enzymes digestion, and then Vpr gene was successfully amplified using its specific primers including restriction sites for XhoI and KpnI. The PCR product was confirmed by electrophoresis. Finally, a recombinant vector pEGFP-N1 containing the full length of human immunodeficiency virus-1 Vpr gene (pEGFP-N1-Vpr) was successfully constructed. Conclusion: HIV-1 Vpr gene in its full length size could be ligated into the pEGFP-N1. [ABSTRACT FROM AUTHOR]

Hemoglobinopathies are most common inherited disorders in the world; approximately 7 percent of the worldwide population and 5-6 percent of population of Iran are carriers. The hemoglobin disorders inherit as autosomal recessive and are very common in the Mediterranean area and much of the Asia and Africa. The control of this inherited disorders need to genetic counseling and accurate screening by more advanced and more accurate methods. This study explains features of current Iran hemoglobin disorders, nominates the accessible methods for screening them and introduces the capillary zone electrophoresis as a rapid and more accurate method. The required data were extracted of various articles and then for good explanation, current Iran hemoglobinopathies properties were showed in the tables and electropherograms of important hemoglobin disorders in Iran population were provided for help to interpretation results of blood tests by capillary zone electrophoresis method. Hemoglobin disorders are including thalassemias and hemoglobin variants; Disruption in the production and malfunction of globin chains cause types of hemoglobin disorders. We cannot introduce one of clinical laboratory tests as critical and basic method for screening and distinguishing types of inherited hemoglobin disorders as alone. For distinguishing the types of them must be prepared enough information and data of the hemoglobin disorders and for more accurate analysis must be used simultaneously different methods as gel electrophoresis, high performance liquid chromatography, isoelectric focusing, capillary zone electrophoresis or molecular tests. The capillary electrophoresis is an accurate and rapid method for screening types of the hemoglobin disorders. Other side this method cannot analyze all of them, so must be used biochemical, biophysical and molecular methods for confirmation the results. This review showed we can use the capillary electrophoresis and HPLC as two complementary methods for hemoglobinopathies screening. We can analyze by the methods more hemoglobin disorders and decrease more laboratory errors. Moreover, we must have patient history, hematological indices, information and data of types of hemoglobinopathies. The patient history and complete blood count results as red blood cell count, mean corpuscular volume, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration can be useful and helpful in screening the hemoglobin disorders and then distinguishing all of hemoglobin disorders. [ABSTRACT FROM AUTHOR]

The country Iran is one of the most important centers for diversity of the genus Verbascum. This study was carried out to determinate intraspecific diversity of the V. speciosum populations collected from East Azerbaijan, West Azerbaijan, Kordestan, Kermanshah and Hamedan provinces. With using the D.S.S. method seventeen special habitats were determined for V. speciosum and floristic-ecologic data collected from each special habitat. Seed storage proteins were subjected for electrophoresis studies. In the survey of all special stations, 109 plant species were distinguished as associated species. As the result of analysis of floristic data, as floristic marker, four groups were determined that this grouping is distinguished the existence of intraspecific diversity in V. speciosum. Analysis of ecologic data was also confirmed the above mentioned grouping. The analysis of seed storage protein banding pattern showed four quite obvious groups. In the survey of electrophoresis pattern, existence of differences regarding number and density of the protein bands indicating existence of intraspecific diversity in the populations of V. speciosum. Therefore in this species, the grouping that introduced with floristic marker, mostly confirmed also with ecological and electrophoresis markers. This means that floristic grouping can only be used as a cost-effective and efficient method for studying of intraspecific diversity. [ABSTRACT FROM AUTHOR]

Background and Objectives Using suitable laboratory methods for the diagnosis of hemoglobinopathies in laboratories not equipped with automatic systems is essential. The correct diagnosis of hemoglobin Q-Iran (α75Asp → His) is important. Case A 33-year-old woman was referred to the national reference globin chain biosynthesis laboratory. The patient's globin chains chromatogram indicated an unknown peak after a globin chain peak. Direct conventional sequencing revealed single G to C missense mutation in the a-globin gene. The genetic analysis led to the identification of a rare hemoglobin variant. Conclusions Medical diagnosis laboratories not equipped with modern automatic systems must run solubility test and hemoglobin electrophoresis on citrate agar and cellulose acetate for definite detection of Hb Q-Iran; otherwise, it can be regarded as a cause of misdiagnosis with hemoglobin S. [ABSTRACT FROM AUTHOR]

Background: SLC26A4 gene mutations are the second identifiable genetic cause of autosomal recessive non-syndromic hearing loss (ARNSHL) after GJB2 mutations. In this study, the identity and informativeness of D7S2459 marker in SLC26A4 gene region was examined in five ethnic groups of the Iranian population. Materials and Methods: The study was accomplished by genotyping the locus in 165 individuals of five different ethnic groups including Fars, Azari, Torkaman, Gilak and Arab using polymerase chain reaction (PCR) followed by polyacrylamide gel electrophoresis (PAGE) and fluorescent capillary electrophoresis. In this study, results were analyzed by GeneMarker HID Human STR Identity software, GenePop program and Microsatellite Tools software. Results: Analysis of the allelic frequency revealed the presence of 8 alleles of D7S2459 marker in the Iranian population. Among all, allele 148bp at the D7S2459 locus was the most frequent with 31% frequency. The most heterozygosity observed in Azari ethnic by 84.8%. Finally, analysis of PIC value revealed that D7S2459 marker could be considered as a highly informative marker in each ethnic of the Iranian population (PIC value above 0.7). Conclusion: Our data suggested that D7S2459 could be introduced as a highly informative marker in molecular diagnosis of SLC26A4 based ARNSHL by Linkage analysis. [ABSTRACT FROM AUTHOR]

Background and Objectives Major p-thalassemia is one of the most problematic diseases both for patients and their families as well as for health services. The number of major thalassemia cases born in 1988 was reported to be 1200 which reduced to 250 in 2007. In spite of the success of the screening program, we still experience some new cases of p-thalassemia. This study aimed to investigate the causes of occurrence of new affected infants to help the prevention program. Materials and Methods In this descriptive analytic study, the parents of p-thalassemia patients having referred were investigated. EDTA blood samples were used for CBC, electrophoresis, and Hb A2 measurment. Simultaneously, the participants were asked to fill out the prepared questionnaire for the causes of occurrence to be evaluated. CBC was performed by sysmex XS800i, electrophoresis by cellulose acetate, and Hb A2 by column chromatography. Results Sixty eight couples, as the parents of 78 patients with major thalassemia, participated in the project. Out of the total number of patients, 20 (25.6%) were under 13 years of age out of whom 7 , 2 and 11 were addressed by the first, second, and third strategies, respectively. Four patients were born due to being left undiagnosed or misdiagnosed due to error in screening and pre-natal tests. Conclusions In order to minimize the occurrence of new cases, we should promote the education and awareness in underdeveloped and non-urban communities. The couples should be identified and directed. The financial support and the possibility of on time testing could be helpful. [ABSTRACT FROM AUTHOR]

Intarspecific and interspecific variations are the main reserves of biodiversity and both are important sources of speciation. On this basis, identifing and recognizing the intra and interspecific variations is important in order to recognition of biodiversity. This research was done to study biodiversity and electrophoresis comparison of seed storage proteins in the populations of the two species of the genus Achillea in Hamadan and Kurdistan provinces using of the method of determination of special station (DSS). For this purpose, 12 and 9 special stations were selected for the species A. tenuifolia and A. biebresteinii using the data published in the related flora. Seed storage proteins were extracted and then studied using electrophoresis techniques (SDS-PAGE). In survey of all special stations, 120 plant species were distinguished as associated species. The results of the floristic data for the both species determined six distinctive groups that indicated the existence of intraspecific diversity in this species. The result of analysis of ecological data and seed storage proteins for the two species was in accordance with the floristic data and showed six distinctive groups. The existence of the bands of no. 4, 5, 8, 12 and 13 in the special stations of A. tenuifolia and the bands of 14, 15 and 16 in the special stations of A. biebresteinii o separated the populations of the species in two quite different and distinctive groups. [ABSTRACT FROM AUTHOR]

Background: The infections caused by Salmonella enterica serovar enteritidis (Salmonella enteritidis) have been increasing in developed and developing countries. The aim of this study was to investigate the genetic relationship between the strains of Salmonella enteritidis. Methods: In a descriptive study from 2008 to 2010, clinical Salmonella enteritidis strains were collected from the patients referred to different hospitals in Tehran. The strains identified by biochemical and serological methods. Genetic relationships between the isolates were investigated using pulsed field gel electrophoresis (PFGE) method. Findings: Of 40 Salmonella enteritidis isolated strains, two PFGE profiles were obtained which among them, 32 (80%) and 8 (20%) of isolates belonged to cluster A and cluster B, respectively. Based on PFGE patterns, 28 (70%) strains exhibited nine bands and were belonged to cluster A. Conclusion: The result of this study indicated that isolated Salmonella enteritidis strains belonged to the closed clones. [ABSTRACT FROM AUTHOR]

Background: Metabolic impacts of two important alleles of the growth hormone receptors (GHR), one with retention of exon 3 [exon 3 (+) GHR] and the other with the deletion of it [exon 3 (-) GHR], on the metabolic risk factors of type 2 diabetes mellitus (T2DM) are really unknown. We aimed to evaluate relationships between the genotypes and metabolic risk factors of T2DM. Methods: In the present study, 40 patients in pre-diabetic state and 40 healthy subjects were selected based on their clinical and laboratory evidence. For genotyping, DNA was extracted from the leukocytes of peripheral blood and amplified by multiplex polymerase chain reaction method. Genotypes of the amplified DNA samples were resolved using 1.5% agarose gel electrophoresis. Other risk factors were also determined by using standard methods and appropriate kits. Findings: Frequency of the homozygote exon 3 retained genotype [exon 3 (+/+) GHR] was significantly higher in the subject in pre-diabetic state (P < 0.001); the frequency of exon 3 (+/-) GHR and exon 3 (-/-) GHR genotypes were significantly higher in the control subjects (P < 0.001). The decrease of metabolic risk factors was profound in subjects with exon 3(-/-) GHR genotype (P < 0.001). Conclusion: The presence of exon 3 deleted allele of GHR, especially in the homozygosity situation, was associated with decreased levels of the metabolic risk factors of type 2 diabetes mellitus. So, the exon 3 deleted GHR may have more robust biological and metabolic activities lead to the resistance against T2DM. [ABSTRACT FROM AUTHOR]

Background: The aim of this study was production of a recombinant antigen from envelope protein of dengue virus, in order to achieve high level expression in soluble form. Therefore, the possibility of envelope protein domain III expression in bacterial host was studied. Methods: Multiple sequence alignment for domain III sequences was carried out using Megalign software. The Modeller and Optimizer software were used for protein structure prediction and sequence optimization. After gene synthesis and subcloning in the pET21a expression vector, the optimization of recombinant protein expression was carried out in Escherichia coli. Ni-NTA chromatography columns were used for protein purification. The efficiency of protein expression was analyzed using electrophoresis and western blotting methods. Findings: A consensus sequence for domain III of dengue virus type 2 envelope protein was provided. The structural properties of target protein were predicted using bioinformatics methods. For high expression level, the sequence of EDIII2 gene was optimized for appropriate codon usage and GC content. The coding gene sequence was approved by sequencing and enzymatic digestion. For disulfide bound formation in protein structure, the Origami (DE3) expression host was used. At the final step, expression of target protein was optimized, and a high concentration of recombinant protein was achieved in soluble form. Conclusion: The results of this study showed that the optimization of domain III sequence led to obtain high concentration (20 mg/l) of produced protein. This expression system can be used for production of dengue virus envelope protein domain III. [ABSTRACT FROM AUTHOR]

Background: Aromatase is a key enzyme in production of estrogen. This enzyme is encoded by cytochrome P-450 (CYP19) gene. It has been shown that overexposure of cells to estrogen causes breast carcinoma and breast cancer patients show higher level of aromatase expression. The purpose of this study was to investigate polymorphism of the CYP19 intron 4 (TTTA)n among breast cancer patients and healthy individuals and its relation to risk of breast cancer. Methods: This study was a case-control study on 100 patients and 100 controls women. DNA was extracted from the blood of study subjects, the sequence were amplified by the technique of Polymerase Chain Reaction (PCR). Thereafter the number and sequence of TTTA was measured by gel electrophoresis on Polyacrylamid. Findings: The results of this study shown that CYP19 gene allele distribution in Isfahan population varies between 7 and 13. The most common allele in both controls and cases was (TTTA)7-TCT with frequencies of 34.3% and 34.8%, respectively. Conclusion: Our results show that the frequency distribution of 10 TTTA repeats is more in controls (2.3%) than in patients (1.3%). This discrepancy suggests a protective role for this allele for Isfahanian women. [ABSTRACT FROM AUTHOR]

Background: Cutaneous Leishmaniasis is endemic in plenty of Iranian provinces. This study aimed to determine the epidemiological status of the cutaneous Leishmaniasis outbreak, isolation and identification of the parasite using a PCR method in burden rural areas of Gonbad-e-Qabus County, north Iran. Methods: Data was collected on the prevalence of scars and ulcers over a period of three months among 6990 inhabitants of five villages around Gonbad-e-Qabus county, north Iran, during 2006-2007. Cultured promastigotes were identified using PCR technique. ITS1 and ITS2 of Non Coding Transcribed region at ribosomal DNA of 46 Leishmania isolates were amplified and the PCR products were separated by electrophoresis in 1.5% agarose gel (200 mA, 140 V), visualized by staining with ethidium bromide, and photographed. To confirm the PCR findings, six Leishmanis isolates were injected individually into two BALB/c mice. Results: Among 6990 inhabitants of the five villages, 62.9% had scars and 0.5% had active lesions. The most highly infected age group was 0-10 years and nobody was infected in individuals more than fifty years of age. Individuals 11 to 20 years of age were the most highly infected age group. The results showed that from 46 isolates, all (100%) were L. major in comparison to reference strains and all of them could produce ulcer at the base tail of BALB/c mice, 4-12 weeks after inoculation. Conclusions: According to this study, cutaneous Leishmaniasis due to Leishmania major is endemic in Gonbad-e-Qabus county, north Iran. The results were confirmed by active lesions induced in BALB/c mice. [ABSTRACT FROM AUTHOR]

Objective: The aim of this study was to investigate the preventive effect of the polyamine component N, N, N′, N′-tetrakis(2-aminoethyl)2,2-dimethylproparie-1,3-diamine (tdmtn) on apoptotic motor neurons in adult mouse spinal cord slices. Materials and Methods: Thoracic region of adult mouse spinal cords was sliced by a tissue chopper into 400 urn slices and cultured in medium in the presence or absence of tdmtn for 6 hours. Morphological a features of apoptosis were evaluated using fluorescent staining with propidium iodide and Hoechst 33342. The appearance of nucleosomal DNA fragmentation was studied using agarose gel electrophoresis. Our results were analyzed using the one-way ANOVA and Tukey's tests. Results: After 6 hours in culture, motor neurons displayed morphological sings of apoptosis including cell shrinkage, as well as nuclear and chromatin condensation. DNA extracted from slices cultured for 24 hours revealed nucleosomal DNA fragmentation on agarose gel electrophoresis. Tdmtn reduced the occurrence of apoptosis in motor neurons and significantly (p<0.01) increased the viability of these neurons after 6 hours of culturing. Conclusion: Tdmtn, as a polyamine component, could probably prevent the occurrence of motor neuron apoptosis through calcium chelation. [ABSTRACT FROM AUTHOR]

Background: Multiple myeloma is a plasma cell dyscrasia characterized by proliferation of plasma cells in bone marrow associated with the production of monoclonal immunoglobulins. In recent years, the use of arsenic trioxide, formerly approved for treatment of acute promyelocytic leukemia has been considered for refractory myeloma treatment. This study was designed and carried out to evaluate the efficacy and possible side effects of ATO on patients with refractory multiple myeloma. Methods: This study carried out on myeloma patients whose diseases were at least refractory to two standard treatment regimens conducted in Ghazi Tabatabaei Hospital in Tabriz- Iran. Arsenic trioxide was administered as an intravenous infusion at a dose of 0.25 mg/kg/d for 5 d/week during the first two consecutive weeks of each 4-week cycle with two week rest. Patients who completed one 4-weak cycle were evaluated for response to treatment. Results: Twelve patients with refractory disease to conventional treatment regimens received arsenic trioxide. The response to the treatment assessed based on the amount of serum proteins electrophoresis of the 10 patients. Stable disease observed in four patients (33%), progressive disease in five patients (41.6%), complete response in one (3.8%) and the remaning two patients could not be assessed for response (because of increased liver enzymes after the first week). One patient completed six cycles. Some adverse events such as: increase liver enzymes and serum creatinine, neutropenia, pruritus, nausea, vomiting, lower extremities edema, and noninfectious diarrhea were observed. Conclusions: The use of arsenic trioxide is promising in treatment of refractory multiple myeloma. [ABSTRACT FROM AUTHOR]

Background: S-layer protects bacteria to the phagocytosis and prohibits the entry of some biomolecules and adhesion molecules to the matrix proteins which increase the stability of infection and antibiotic resistance in virulence agents. Therefore the spread of bacteria which have S-layer in the hospitals should be avoided. Surfaces can serve as reservoirs of potential pathogen and is important in the chain of infection. Surfaces bacteria have low potential to spread, but staff hands have many contacts with hospital surfaces, especially with high contact surfaces. Therefore it could be a potential source of bacteria transmission from surfaces. Methods: This research was performed in the Al-zahra hospital of Isfahan University of Medical Sciences (2005 to 2007). 274 samples were randomly selected (194 sample from surface and 80 sample from staff hand). Environmental samples collected, from high and low contact surface with swab in NB and staff hand. Samples were collected through finger print method and identification of bacteria, were performed with microbiological methods: staining, chemical test, use of differential and selective media. Isolated Bacillus cereus strains, performed with Bacillus cereus Selective Agar, for preparation samples, in first culture Bacteria in TSA, for 16h, then separated surface proteins and finally, specimen's electrophoresis proximally molecular weight marker. S-layer in Bacillus cereus has 97KD molecular weight. Findings: According to the results, the prevalence of Bacillus cereus strains was 9.49%. The prevalence of Bacillus cereus strains in environmental and staff hands were respectively 6.7% and 16.25%, and of 13 isolated Bacillus cereus from, staff hand 11 strain (84.6%) and from 13 isolated Bacillus cereus from environmental samples 1 strain (7.7%) production S-layer. Conclusion: We found a high prevalence Bacillus cereus in staff hands and hospital surfaces and frequency surface layer in Bacillus cereus isolated of staff hands into hospital surfaces. [ABSTRACT FROM AUTHOR]